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Updated: Sep 26, 2025
Generation of Retinal Organoids from Healthy and Retinal Disease-Specific Human-Induced Pluripotent Stem Cells
Published on: December 9, 2022
Characterization of The Retinal Progenitor Cells Generated Using Co-Culture Systems
Sara Momenzadeh1,2, Fereshteh Karamali2, Atefeh Atefi2
1Higher Education Jahad University of Isfahan Province, Isfahan, Iran.
Objective:
Degeneration of the photoreceptors due to retinal disorders can affect vision, and even lead to blindness. Recently therapeutic progress in retinal degeneration, using human embryonic stem cells (hESCs), has been facing technical challenges, demanding the development of simple and standardized protocols. In addition to the designing of the protocols, characterization of the obtained cells is highly required for confirming the reliability of the applied methods for future medical applications. Previously, we showed that human stem cells from apical papilla (SCAP) have stromal cell-derived inducing activity (SDIA).
Materials And Methods:
In this experimental study, we developed an efficient retinal differentiation protocol, based on the co-culture of confluent hESCs and SCAP in the absence of exogenous molecules, such as activators or inhibitors of molecular signaling pathways. This experimental procedure resulted in the generation of self-forming neural retina (NR)-like structures containing retinal progenitor cells (RPCs) within 4 weeks.
Results:
We have focused on the characterization of the derived RPCs, as a crucial step towards further verification of the efficiency of our previously suggested protocol. The differentiated cells expressed eye-field markers, PAX6, RAX, LHX2, and SIX3, and also generated neurospheres by a floating culture system for one week.
Conclusion:
We have reported that the treatment of hESC-derived RPCs by the Notch pathway-inhibitor induced the generation of photoreceptor precursor cells (PPCs). The presented method demonstrates the fact that a co-culture of hESCs and SCAP without exogenous molecules provides an efficient approach to produce RPCs for the treatment of retinal disease, and act as an in vitro model for the development of human retina.
Insights
This study developed a simple protocol using human embryonic stem cells (hESCs) and stem cells from apical papilla (SCAP) to generate retinal progenitor cells (RPCs). This method offers a new avenue for treating retinal degeneration and developing human retina models.
Area of Science:
- Stem cell biology
- Ophthalmology
- Regenerative medicine
Background:
- Retinal degeneration leads to vision loss and blindness.
- Current therapeutic strategies using human embryonic stem cells (hESCs) face technical hurdles.
- Standardized protocols and cell characterization are crucial for reliable retinal therapies.
Purpose of the Study:
- To develop an efficient and standardized protocol for retinal differentiation.
- To characterize the generated retinal progenitor cells (RPCs).
- To establish a co-culture system for generating retinal cells without exogenous molecules.
Main Methods:
- Co-culture of confluent hESCs and SCAP without signaling pathway modulators.
- Generation of self-forming neural retina (NR)-like structures.
- Characterization of derived RPCs using eye-field markers and neurosphere formation.
Main Results:
- Efficient generation of NR-like structures containing RPCs within 4 weeks.
- Differentiated cells expressed key eye-field markers (PAX6, RAX, LHX2, SIX3).
- RPCs formed neurospheres in a floating culture system.
Conclusions:
- Co-culture of hESCs and SCAP provides an efficient method for producing RPCs.
- This approach bypasses the need for exogenous molecules, simplifying the protocol.
- The generated RPCs can be used for retinal disease treatment and as an in vitro human retina model.
